1. Neuroscience
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Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry

  1. Seksiri Arttamangkul  Is a corresponding author
  2. Andrew Plazek
  3. Emily J Platt
  4. Haihong Jin
  5. Thomas F Murray
  6. William Birdsong
  7. Kenner C Rice
  8. David Farrens
  9. John T Williams
  1. Oregon Health and Science University, United States
  2. Creighton University, United States
  3. National Institute on Drug Abuse/National Institute on Alcohol Abuse and Alcoholism, United States
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Cite this article as: eLife 2019;8:e49319 doi: 10.7554/eLife.49319

Abstract

Identifying neurons that have functional opioid receptors is fundamental for the understanding of the cellular, synaptic and systems actions of opioids. Current techniques are limited to post hoc analyses of fixed tissues. Here we developed a fluorescent probe, naltrexamine-acylimidazole (NAI), to label opioid receptors based on a chemical approach termed 'traceless affinity labeling'. In this approach, a high affinity antagonist naltrexamine is used as the guide molecule for a transferring reaction of acylimidazole at the receptor. This reaction generates a fluorescent dye covalently linked to the receptor while naltrexamine is liberated and leaves the binding site. The labeling induced by this reagent allowed visualization of opioid-sensitive neurons in rat and mouse brains without loss of function of the fluorescently labeled receptors. The ability to locate endogenous receptors in living tissues will aid considerably in establishing the distribution and physiological role of opioid receptors in the CNS of wild type animals.

Data availability

All data generated or analysed during this study are includes in the manuscript and supporting files.

Article and author information

Author details

  1. Seksiri Arttamangkul

    Vollum Institute, Oregon Health and Science University, Portland, United States
    For correspondence
    arttaman@ohsu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8815-5124
  2. Andrew Plazek

    Medicinal Chemistry Core, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Emily J Platt

    Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Haihong Jin

    Medicinal Chemistry Core, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Thomas F Murray

    Department of Pharmacology, School of Medicine, Creighton University, Omaha, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. William Birdsong

    Vollum Institute, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Kenner C Rice

    Drug Design and Synthesis Section, Intramural Research Program, National Institute on Drug Abuse/National Institute on Alcohol Abuse and Alcoholism, Bethesda, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. David Farrens

    Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. John T Williams

    Vollum Institute, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0647-6144

Funding

National Institute on Drug Abuse (DA008163)

  • John T Williams

National Institute on Drug Abuse (DA048136)

  • John T Williams

National Institute on Drug Abuse (DA042779)

  • William Birdsong

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal uses were conducted in accordance with the National Institutes of Health guidelines and with approval from the Institutional Animal Care and Use Committee (IACUC) protocol #IP00000160 of the Oregon Health & Science University. Rats and mice were anesthetized with isofluorane before euthanized with minimal suffering.

Reviewing Editor

  1. Peggy Mason, University of Chicago, United States

Publication history

  1. Received: July 4, 2019
  2. Accepted: October 6, 2019
  3. Accepted Manuscript published: October 7, 2019 (version 1)
  4. Version of Record published: October 23, 2019 (version 2)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

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